This invention concerns certain amide derivatives and their use as inhibitors of cytokine mediated disease. The invention also concerns processes for the manufacture of said novel amide derivatives, pharmaceutical compositions containing them and their use in therapeutic methods, for example by virtue of inhibition of cytokine mediated disease.
The amide derivatives disclosed in the present invention are inhibitors of the production of cytokines such as Tumour Necrosis Factor (hereinafter TNF), for example TNFxcex1, and various members of the interleukin (hereinafter IL) family, for example IL-1, IL-6 and IL-8. Accordingly the compounds of the invention will be useful in the treatment of diseases or medical conditions in which excessive production of cytokines occurs, for example excessive production of TNFxcex1 or IL-1. It is known that cytokines are produced by a wide variety of cells such as monocytes and macrophages and that they give rise to a variety of physiological effects which are believed to be important in disease or medical conditions such as inflammation and immunoregulation. For example, TNFxcex1 and IL-1 have been implicated in the cell signalling cascade which is believed to contribute to the pathology of disease states such as inflammatory and allergic diseases and cytokine-induced toxicity. It is also known that, in certain cellular systems, TNFxcex1 production precedes and mediates the production of other cytokines such as IL-1.
Abnormal levels of cytokines have also been implicated in, for example, the production of physiologically-active eicosanoids such as the prostaglandins and leukotrienes the stimulation of the release of proteolytic enzymes such as collagenase, the activation of the immune system, for example by stimulation of T-helper cells, the activation of osteoclast activity leading to the resorption of calcium, the stimulation of the release of proteoglycans from, for example, cartilage, the stimulation of cell proliferation and to angiogenesis.
Cytokines are also believed to be implicated in the production and development of disease states such as inflammatory and allergic diseases, for example inflammation of the joints (especially rheumatoid arthritis, osteoarthritis and gout), inflammation of the gastrointestinal tract (especially inflammatory bowel disease, ulcerative colitis, Crohn""s disease and gastritis), skin disease (especially psoriasis, eczema and dermatitis) and respiratory disease (especially asthma, bronchitis, allergic rhinitis and adult respiratory distress syndrome), and in the production and development of various cardiovascular and cerebrovascular disorders such as congestive heart disease, myocardial infarction, the formation of atherosclerotic plaques, hypertension, platelet aggregation, angina, stroke, reperfusion injury, vascular injury including restenosis and peripheral vascular disease, and, for example, various disorders of bone metabolism such as osteoporosis (including senile and postmenopausal osteoporosis), Paget""s disease, bone metastases, hypercalcaemia, hyperparathyroidism, osteosclerosis, osteoporosis and periodontitis, and the abnormal changes in bone metabolism which may accompany rheumatoid arthritis and osteoarthritis. Excessive cytokine production has also been implicated in mediating certain complications of bacterial, fungal and/or viral infections such as endotoxic shock, septic shock and toxic shock syndrome and in mediating certain complications of CNS surgery or injury such as neurotrauma and ischaemic stroke. Excessive cytokine production has also been implicated in mediating or exacerbating the development of diseases involving cartilage or muscle resorption, pulmonary fibrosis, cirrhosis, renal fibrosis, the cachexia found in certain chronic diseases such as malignant disease and acquired immune deficiency syndrome (AIDS), tumour invasiveness and tumour metastasis and multiple sclerosis.
Evidence of the central role played by TNFxcex1 in the cell signalling cascade which gives rise to rheumatoid arthritis is provided by the efficacy in clinical studies of antibodies of TNFxcex1 (The Lancet, 1994, 344, 1125 and British Journal of Rheumatology, 1995, 34, 334).
Thus cytokines such as TNFxcex1 and IL-1 are believed to be important mediators of a considerable range of diseases and medical conditions. Accordingly it is expected that inhibition of the production of and/or effects of these cytokines will be of benefit in the prophylaxis, control or treatment of such diseases and medical conditions.
Without wishing to imply that the compounds disclosed in the present invention possess pharmacological activity only by virtue of an effect on a single biological process, it is believed that the compounds inhibit the effects of cytokines by virtue of inhibition of the enzyme p38 kinase. p38 kinase, otherwise known as cytokine suppressive binding protein (hereinafter CSBP) and reactivating kinase (hereinafter RK), is a member of the mitogen-activated protein (hereinafter MAP) kinase family of enzymes which is known to be activated by physiological stress such as that induced by ionising radiation, cytotoxic agents, and toxins for example endotoxins such as bacterial lipopolysaccharide, and by a variety of agents such as the cytokines, for example TNFxcex1 and IL-1. It is known that p38 kinase phosphorylates certain intracellular proteins which are involved in the cascade of enzymatic steps which leads to the biosynthesis and excretion of cytokines such as TNFxcex1 and IL-1. Known inhibitors of p38 kinase have been reviewed by G J Hanson in Expert Opinions on Therapeutic Patents. 1997 7, 729-733. p38 kinase is known to exist in isoforms identified as p38xcex1 and p38xcex2.
European Patent Application No. 0 566 226, discloses certain quinazoline compounds as tyrosine kinase-inhibiting anticancer agents including the compounds:
4-(3-acetamidoanilino)-6,7-dimethoxyquinazoline and
4-(3-benzamidoanilino)-6,7-dimethoxyquinazoline.
The compounds disclosed in the present invention are inhibitors of the production of cytokines such as TNF, in particular of TNFxcex1, and various interleukins, in particular IL-1.
According to one aspect of the present invention there is provided a compound of the Formula (I): 
wherein:
G is N or CH;
R1 is hydroxy, halo, trifluoromethyl, cyano, mercapto, nitro, amino, carboxy, carbamoyl, formyl, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, xe2x80x94O-(C1-3alkyl)-Oxe2x80x94, C1-6alkylS(O)n- (wherein n is 0-2), N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, C1-6alkoxycarbonyl, N-C1-6alkylcarbamoyl, N,N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, C1-6alkanoyloxy, C1-6alkanoylamino, N-C1-6alkylsulphamoyl, N,N-(C1-6alkyl)2sulphamoyl, C1-6alkylsulphonylamino, C1-6alkylsulphonyl-N-(C1-6alkyl)amino, or R1 is of the Formula (IA):
Axe2x80x94(CH2)pxe2x80x94Bxe2x80x94xe2x80x83xe2x80x83(IA)
wherein A is halo, hydroxy, C1-6alkoxy, C1-6alkylS(O)n- (wherein n is 0-2), cyano, amino, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, carboxy, C1-6alkoxycarbonyl, carbamoyl, N-C1-6alkylcarbamoyl or N,N-(C1-6alkyl)2carbamoyl, p is 1-6, and B is a bond, oxy, imino, N-(C1-6alkyl)imino or xe2x80x94C(O)NHxe2x80x94, with the proviso that p is 2 or more unless B is a bond or xe2x80x94C(O)NHxe2x80x94,
or R1 is of the Formula (IB):
Dxe2x80x94Exe2x80x94xe2x80x83xe2x80x83(IB)
wherein D is aryl, heteroaryl or heterocyclyl and E is a bond, C1-6alkylene, C1-6alkeneoxy, oxy, imino, N-(C1-6alkyl)imino, C1-6alkyleneimino, N-(C1-6alkyl)-C1-6alkyleneimino, C1-6alkyleneoxy-C1-6alkylene, C1-6alkyleneimino-C1-6alkylene, N-(C1-6alkyl)-C1-6alkyleneimino-C1-6alkylene, xe2x80x94C(O)NHxe2x80x94, xe2x80x94SO2NHxe2x80x94, xe2x80x94NHSO2xe2x80x94 or C2-6alkanoylimino, and any aryl, heteroaryl or heterocyclyl group in a R1 group may be optionally substituted with one or more groups selected from hydroxy, halo, C1-6alkyl, C1-6alkoxy, carboxy, C1-6alkoxycarbonyl, carbamoyl, N-C1-6alkylcarbamoyl, N,N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, amino, N-C1-6alkylamino and N,N-(C1-6alkyl)2amino,
and any heterocyclyl group in a R1 group may be optionally substituted with one or two oxo or thioxo substituents,
and any of the R1 groups defined hereinbefore which comprises a CH2 group which is attached to 2 carbon atoms or a CH3 group which is attached to a carbon atom may optionally bear on each said CH2 or CH3 group a substituent selected from hydroxy, amino, C1-6alkoxy,
N-C1-6alkylamino, N,N-(C1-6alkyl)2amino and heterocyclyl;
R2 is hydrogen, halo, C1-6alkyl, C2-6alkenyl or C2-6alkynyl;
R3 is hydrogen, halo, C1-6alkyl, C2-6alkenyl or C2-6alkynyl;
R4 is hydrogen, hydroxy, C1-6alkyl, C1-6alkoxy, amino, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, hydroxyC2-6alkoxy, C1-6alkoxyC2-6alkoxy, aminoC2-6alkoxy, N-C1-6alkylaminoC2-6alkoxy, N,N-(C1-6alkyl)2aminoC2-6alkoxy or C3-7cycloalkyl,
or R4 is of the Formula (IC):
xe2x80x94Kxe2x80x94Jxe2x80x83xe2x80x83(IC)
wherein J is aryl, heteroaryl or heterocyclyl and K is a bond, oxy, imino, N-(C1-6alkyl)imino, oxyC1-6alkylene, iminoC1-6alkylene, N-(C1-6alkyl)iminoC1-6alkylene, xe2x80x94NHC(O)xe2x80x94, xe2x80x94SO2NHxe2x80x94, xe2x80x94NHSO2xe2x80x94 or xe2x80x94NHC(O)xe2x80x94C1-6alkylene-,
and any aryl, heteroaryl or heterocyclyl group in a R4 group may be optionally substituted by one or more groups selected from hydroxy, halo, trifluoromethyl, cyano, mercapto, nitro, amino carboxy, carbamoyl, formyl, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, xe2x80x94O-(C1-3alkyl)-Oxe2x80x94, C1-6alkylS(O)nxe2x80x94 (wherein n is 0-2), N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, C1-6alkoxycarbonyl, N-C1-6alkylcarbamoyl, N,N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, C1-6alkanoyloxy, C1-6alkanoylamino, N-C1-6alkylsulphamoyl, N,N-(C1-6alkyl)2sulphamoyl, C1-6alkylsulphonylamino and C1-6alkylsulphonyl-N-(C1-6alkyl)amino,
or any aryl, heteroaryl or heterocyclyl group in a R4 group may be optionally substituted with one or more groups of the Formula (IAxe2x80x2):
xe2x80x94B1xe2x80x94(CH2)pxe2x80x94A1xe2x80x83xe2x80x83(IAxe2x80x2)
wherein A1 is halo, hydroxy, C1-6alkoxy, cyano, amino, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, carboxy, C1-6alkoxycarbonyl, carbamoyl, N-C1-6alkylcarbamoyl or N,N-(C1-6alkyl)2carbamoyl, p is 1-6, and B1 is a bond, oxy, imino, N-(C1-6alkyl)imino or xe2x80x94NHC(O)xe2x80x94, with the proviso that p is 2 or more unless B1 is a bond or xe2x80x94NHC(O)xe2x80x94;
or any aryl, heteroaryl or heterocyclyl group in a R4 group may be optionally substituted with one or more groups of the Formula (IBxe2x80x2):
xe2x80x94E1xe2x80x94D1xe2x80x83xe2x80x83(IBxe2x80x2)
wherein D1 is aryl, heteroaryl or heterocyclyl and E1 is a bond, C1-6alkylene, oxyC1-6alkylene, oxy, imino, N-(C1-6alkyl)imino, iminoC1-6alkylene, N-(C1-6alkyl)iminoC1-6alkylene, C1-6alkylene-oxyC1-6alkylene, C1-6alkylene-iminoC1-6alkylene, C1-6alkylene-N-(C1-6alkyl)-iminoC1-6alkylene, xe2x80x94NHC(O)xe2x80x94, xe2x80x94NHSO2xe2x80x94, xe2x80x94SO2NHxe2x80x94 or xe2x80x94NHC(O)xe2x80x94C1-6alkylene-, and any aryl, heteroaryl or heterocyclyl group in a substituent on R4 may be optionally substituted with one or more groups selected from hydroxy, halo, C1-6alkyl, C1-6alkoxy, carboxy, C1-6alkoxycarbonyl, carbamoyl, N-C1-6alkylcarbamoyl, N,N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, amino, N-C1-6alkylamino and N,N-(C1-6alkyl)2amino,
and any C3-7cycloalkyl or heterocyclyl group in a R4 group may be optionally substituted with one or two oxo or thioxo substituents,
and any of the R4 groups defined hereinbefore which comprises a CH2 group which is attached to 2 carbon atoms or a CH3 group which is attached to a carbon atom may optionally bear on each said CH2 or CH3 group a substituent selected from hydroxy, amino, C1-6alkoxy, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino and heterocyclyl;
R5 is hydrogen, halo, trifluoromethyl, cyano, nitro, amino, hydroxy, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, N-C1-6alkylamino or N,N-(C1-6alkyl)2amino;
m is 1, 2 or 3; and
q is 0, 1, 2, 3 or 4;
or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof; with the proviso that:
4-(3-acetamidoanilino)-6,7-dimethoxyquinazoline; and
4-(3-benzamidoanilino)-6,7-dimethoxyquinazoline are excluded.
According to a further aspect of the present invention there is provided a compound of the Formula (I) wherein:
G is N or CH;
R1 is hydroxy, halo, trifluoromethyl, cyano, mercapto, nitro, amino, carboxy, carbamoyl, formyl, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, xe2x80x94O-(C1-3alkyl)-Oxe2x80x94, C1-6alkylS(O)nxe2x80x94 (wherein n is 0-2), N-C1-6alkylamino, N,N-(C1-4alkyl)2amino, C1-6alkoxycarbonyl, N-C1-6alkylcarbamoyl, N,N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, C1-6alkanoyloxy, C1-6alkanoylamino, N-C1-6alkylsulphamoyl, N,N-(C1-6alkyl)2sulphamoyl, C1-6alkylsulphonylamino, C1-6alkylsulphonyl-N-(C1-6alkyl)amino, or R1 is of the formula:
Axe2x80x94(CH2)pxe2x80x94Bxe2x80x94xe2x80x83xe2x80x83(IA)
wherein A is halo, hydroxy, C1-6alkoxy, cyano, amino, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, carboxy, C1-6alkoxycarbonyl, carbamoyl, N-C1-6alkylcarbamoyl or N,N-(C1-6alkyl)2carbamoyl, p is 1-6, and B is a bond, oxy, imino, N-(C1-6alkyl)imino or xe2x80x94C(O)NHxe2x80x94, with the proviso that p is 2 or more unless B is a bond or xe2x80x94C(O)NHxe2x80x94, or R1 is of the formula:
Dxe2x80x94Exe2x80x94xe2x80x83xe2x80x83(IB)
wherein D is aryl, heteroaryl or heterocyclyl and E is a bond, C1-6alkylene, C1-6alkyleneoxy, oxy, imino, N-(C1-6alkyl)imino, C1-6alkyleneimino, N-(C1-6alkyl)-C1-6alkyleneimino, xe2x80x94C(O)NHxe2x80x94, xe2x80x94SO2NHxe2x80x94, xe2x80x94NHSO2xe2x80x94 or C2-6alkanoylimino, and any aryl, heteroaryl or heterocyclyl group may be optionally substituted with one or more groups selected from hydroxy, halo, C1-6alkyl, C1-6alkoxy, carboxy, C1-6alkoxycarbonyl, carbamoyl, N-C1-6alkylcarbamoyl, N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, amino, N-C1-6alkylamino and N,N-(C1-6alkyl)2amino;
R2 is hydrogen, halo, C1-6alkyl, C2-6alkenyl or C2-6alkynyl;
R3 is hydrogen, halo, C1-6alkyl, C2-6alkenyl or C2-6alkynyl;
R4 is hydrogen, hydroxy, C1-6alkyl, C1-6alkoxy, amino, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, hydroxyC2-6alkoxy, C1-6alkoxyC2-6alkoxy, aminoC2-6alkoxy, N-C1-6alkylaminoC2-6alkoxy, N,N-(C1-6alkyl)2aminoC2-6alkoxy or C3-7cycloalkyl, or R4 is of the formula:
xe2x80x94Kxe2x80x94Jxe2x80x83xe2x80x83(IC)
wherein J is aryl, heteroaryl or heterocyclyl and K is a bond, oxy, imino, N-(C1-6alkyl)imino, oxyC1-6alkylene, iminoC1-6alkylene, N-(C1-6alkyl)iminoC1-6alkylene, xe2x80x94NHC(O)xe2x80x94, xe2x80x94SO2NHxe2x80x94, xe2x80x94NHSO2xe2x80x94 or xe2x80x94NHC(O)xe2x80x94C1-6alkylene-, and any aryl, heteroaryl or heterocyclyl group may be optionally substituted by one or more groups selected from hydroxy, halo, trifluoromethyl, cyano, mercapto, nitro, amino, carboxy, carbamoyl, formyl, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, xe2x80x94O-(C1-3alkyl)-Oxe2x80x94, C1-6alkylS(O)nxe2x80x94 (wherein n is 0-2), N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, C1-6alkoxycarbonyl, N-C1-6alkylcarbamoyl, N,N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, C1-6alkanoyloxy, C1-6alkanoylamino, N-C1-6alkylsulphamoyl, N,N-(C1-6alkyl)2sulphamoyl, C1-6alkylsulphonylamino and C1-6alkylsulphonyl-N-(C1-6alkyl)amino, or any aryl, heteroaryl or heterocyclyl may be optionally substituted with one or more groups of the formula (IAxe2x80x2):
xe2x80x94B1-(CH2)pxe2x80x94A1xe2x80x83xe2x80x83(IAxe2x80x2)
wherein A1 is halo, hydroxy, C1-6alkoxy, cyano, amino, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, carboxy, C1-6alkoxycarbonyl, carbamoyl, N-C1-6alkylcarbamoyl or N,N-(C1-6alkyl)2carbamoyl, p is 1-6, and B1 is a bond, oxy, imino, N-(C1-6alkyl)imino or xe2x80x94NHC(O)xe2x80x94, with the proviso that p is 2 or more unless B1 is a bond or xe2x80x94NHC(O)xe2x80x94; and/or (IBxe2x80x2):
xe2x80x94E1xe2x80x94D1xe2x80x83xe2x80x83(IBxe2x80x2)
wherein D1 is aryl, heteroaryl or heterocyclyl and E1 is a bond, C1-6alkylene, oxyC1-6alkylene, oxy, imino, N-(C1-6alkyl)imino, iminoC1-6alkylene, N-(C1-6alkyl)iminoC1-6alkylene, xe2x80x94NHC(O)xe2x80x94, xe2x80x94NHSO2xe2x80x94, xe2x80x94SO2NHxe2x80x94 or xe2x80x94NHC(O)xe2x80x94C1-6alkylene-, and any aryl, heteroaryl or heterocyclyl group may be optionally substituted with one or more groups selected from hydroxy, halo, C1-6alkyl, C1-6alkoxy, carboxy, C1-6alkoxycarbonyl, carbamoyl, N-C1-6alkylcarbamoyl, N,N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, amino, N-C1-6alkylamino and N,N-(C1-6alkyl)2amino;
R5 is hydrogen, halo, trifluoromethyl, cyano, nitro, amino, hydroxy, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, N-C1-6alkylamino or N,N-(C1-6alkyl)2amino;
m is 1, 2 or 3;
q is 0, 1, 2, 3 or 4;
or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof; with the proviso that:
4-(3-acetamidoanilino)-6,7-dimethoxyquinazoline; and
4-(3-benzamidoanilino)-6,7-dimethoxyquinazoline are excluded.
In this specification the term xe2x80x9calkylxe2x80x9d includes both straight and branched chain alkyl groups but references to individual alkyl groups such as xe2x80x9cpropylxe2x80x9d are specific for the straight chain version only. For example, xe2x80x9cC1-6alkylxe2x80x9d includes propyl, isopropyl and tert-butyl. However, references to individual alkyl groups such as xe2x80x98propylxe2x80x99 are specific for the straight chained version only and references to individual branched chain alkyl groups such as xe2x80x98isopropylxe2x80x99 are specific for the branched chain version only. A similar convention applies to other radicals, for example xe2x80x9caminoC2-6alkoxyxe2x80x9d includes 2-aminoethoxy, 2-aminopropoxy and 3-amino-2-methylpropoxy. The term xe2x80x9chaloxe2x80x9d refers to fluoro, chloro, bromo and iodo.
The term xe2x80x9carylxe2x80x9d refers to phenyl or naphthyl.
The term xe2x80x9cheteroarylxe2x80x9d refers to, unless otherwise further specified, a monocyclic-, bicyclic- or tricyclic-5-14 membered ring that contains some degree of unsaturation, with up to five ring heteroatoms selected from nitrogen, oxygen and sulphur wherein a xe2x80x94CH2xe2x80x94 group can optionally be replaced by a xe2x80x94C(O)xe2x80x94, a ring nitrogen atom may optionally bear a C1-6alkyl group or a ring nitrogen atom may be optionally oxidised to form the N-oxide. Examples of xe2x80x9cheteroarytxe2x80x9d include thienyl, furyl, pyranyl, pyrrolyl, imidazolyl, pyrazolyl, thiazolyl, oxazolyl, isoxazolyl, pyridyl, pyridyl-N-oxide, oxopyridyl, oxoquinolyl, pyrimidinyl, pyrazinyl, oxopyrazinyl, pyridazinyl, indolyl, benzofuranyl, benzimidazolyl, benzothiazolyl, quinolyl, N-methyloxoquinolyl, isoquinolinyl, quinazolinyl, xanthenyl, quinoxalinyl, indazolyl, benzofuranyl and cinnolinolyl.
The term xe2x80x9cheterocyclylxe2x80x9d refers to, unless otherwise further specified, a mono- or bicyclic-5-14 membered ring, that is totally saturated, with up to five ring heteroatoms selected from nitrogen, oxygen and sulphur wherein a xe2x80x94CH2xe2x80x94 group can optionally be replaced by a xe2x80x94C(O)xe2x80x94 or a ring nitrogen atom may optionally bear a C1-6alkyl group. Examples of such heterocyclyls include morpholinyl, N-methylmorpholinyl, pyrrolidinyl, imidazolidinyl, pyrazolidinyl, piperidinyl, homopiperidinyl, N-methylpiperidinyl, piperazinyl, homopiperazinyl and quinuclidinyl.
Where optional substituents are chosen from xe2x80x9cone or morexe2x80x9d groups it is to be understood that this definition includes all substituents being chosen from one of the specified groups or the substituents being chosen from two or more of the specified groups. Conveniently there may be 1, 2 or 3 such optional substituents. For example, where optional substituents are chosen from one or more groups selected from halo, C1-6alkoxy and C1-6alkyl, examples of possible combinations of substituents include 1) a bromo group, 2) two chloro groups, 3) a methoxy, ethoxy and propoxy substituent, 4) a fluoro and a methoxy group, 5) a methoxy, a methyl and an ethyl group, and 6) a chloro, a methoxy and an ethyl group.
It is to be understood that the bicyclic ring within the compound of Formula (I) is shown with a hydrogen atom attached to the carbon between the N atom and G group in order to indicate that this position is unsubstituted. Thereby it is to be understood that that hydrogen atom may not be replaced by a R1 substituent. It should also be understood however that when G is a CH group such that the bicyclic ring is a quinoline ring the 3-position of the quinoline ring may bear any one of the R1 substituents.
The following table gives examples of radicals that fall within the definition of the generic terms used in this specification:
In the linking groups B, E, B1, E1 and K that fall within the definition of R1 and R4, the following table gives examples of radicals that fall within these general terms:
For the avoidance of doubt, it is to be understood that when, for example, R1 is a group of the Formula (IB):
Dxe2x80x94Exe2x80x94xe2x80x83xe2x80x83(IB)
and the linking group E is, for example, a C1-6alkyleneoxy group such as xe2x80x94CH2CH2Oxe2x80x94, it is a CH2 group which is attached to D and the O atom which is attached to the bicyclic ring within Formula (I). Similarly when, for example, R4 is a group of the Formula (IBxe2x80x2):
xe2x80x94E1xe2x80x94D1xe2x80x83xe2x80x83(IBxe2x80x2)
and the linking group E1 is, for example, an iminoC1-6alkylene group such as xe2x80x94NHCH2CH2xe2x80x94, it is a CH2 group which is attached to D1 and the NH group which is attached to the bicyclic ring within Formula (I). An analogous convention applies to other bidentate linking groups.
It is to be understood that, insofar as certain of the compounds of the Formula (I) defined above may exist in optically active or racemic forms by virtue of one or more asymmetric carbon atoms, the invention includes in its definition any such optically active or racemic form which possesses the property of inhibiting cytokines, in particular TNF. The synthesis of optically active forms may be carried out by standard techniques of organic chemistry well known in the art, for example by synthesis from optically active starting materials or by resolution of a racemic form. Similarly, inhibitory properties against TNF may be evaluated using the standard laboratory techniques referred to hereinafter.
Preferable values of R1, R2, R3, R4, R5, G, q and m are as follows.
Preferably R1 is hydroxy, halo, C1-6alkoxy, N,N-(C1-6allyl)2aminoC1-6alkyl, N,N-(C1-6alkyl)2carbamoylC1-6alkoxy, N,N-(C1-6alkyl)2aminoC1-6alkoxy, C1-6alkylS(O)2xe2x80x94C1-6alkoxy, N,N-(C1-6alkyl)2amino-N-(C1-6alkyl)C1-6alkylamino, N,N-(C1-6alkyl)2aminoC1-6alkylaminoC1-6alkyl, heterocyclylC1-6alkyl, heterocyclylC1-6alkoxy, heterocyclyloxy, heterocyclylC1-6alkylaminoC1-6alkyl or heteroarylC1-6alkoxy.
More preferably R1 is hydroxy, halo, C1-6alkoxy, N,N-(C1-6alkyl)2aminoC1-6alkyl, N,N-(C1-6alkyl)2carbamoylC1-6alkoxy, N,N-(C1-6alkyl)2aminoC1-6alkoxy, C1-6alkylS(O)2xe2x80x94C1-6alkoxy, N,N-(C1-6alkyl)2amino-N-(C1-6alkyl)C6alkylamino, N,N-(C1-6alkyl)2aminoC1-6alkylaminoC1-6alkyl, piperazin-1-ylC1-6alkyl, 4-C1-6alkylpiperazin-1-ylC1-6alkyl, homopiperazinyl-1-ylC1-6alkyl, 4-C1-6alkylhomopiperazinyl-1-ylC1-6alkyl, pyrrolidinylC1-6alkoxy, piperidinylC1-6alkoxy, N-(C1-6alkyl)pyrrolidinylC1-6alkoxy, N-(C1-6alkyl)piperidinylC1-6alkoxy, morpholinylC1-6alkoxy, piperazinylC1-6alkoxy, N-(C1-6alkyl)piperazinylC1-6alkoxy, homopiperazinylC1-6alkoxy, N-(C1-6alkyl)homopiperazinyl C1-6alkoxy, pyrrolidinyloxy, piperidinyloxy, morpholinylC1-6alkylaminoC1-6alkyl or pyridylC1-6alkoxy.
More particularly R1 is methoxy, 2-dimethylaminoethoxy, 2-diethylaminoethoxy, 2-diisopropylaminoethoxy, 3-dimethylaminopropoxy, 3-diethylaminopropoxy, 2-morpholinoethoxy, 3-morpholinopropoxy, 2-piperidinoethoxy, N-methylpiperidin-2-ylmethoxy, N-methylpiperidin-3-ylmethoxy, 2-pyrrolidin-1-ylethoxy, 2-(N-methylpyrrolidin-2-yl)ethoxy, N-methyl-5-oxopyrrolidin-2-ylmethoxy, 3-pyrrolidin-1-ylpropoxy, 2-(2-oxoimidazolidin-1-yl)ethoxy, 2-(4-methylpiperazin-1-yl)ethoxy or 3-pyrid-3-ylpropoxy.
Further more particularly R1 is methoxy, 2-diisopropylaminoethoxy, 3-diethylaminopropoxy, 3-morpholinopropoxy or 3-pyrrolidin-1-ylpropoxy.
More preferably R1 is C1-6alkoxy, heterocyclylC1-6alkoxy or heteroarylC1-6alkoxy.
More preferably R1 is C1-6alkoxy, morpholinylC1-6alkoxy, pyrrolidinylC1-6alkoxy, pyridylC1-6alkoxy, piperidin-1-ylC1-6alkoxy, piperazin-1-ylC1-6alkoxy or 4-C1-6alkylpiperazinyl-1-ylC1-6alkoxy.
Particularly R1 is C1-4alkoxy, morpholinylC2-4alkoxy, pyrrolidinylC2-4alkoxy or pyridylC2-4alkoxy.
More particularly R1 is methoxy, 2-morpholinoethoxy, 3-morpholinopropoxy, 2-pyrrolidin-1-ylethoxy or 3-pyrid-3-ylpropoxy.
Preferably R2 is hydrogen, C1-6alkyl or halo.
More preferably R2 is hydrogen, C1-4alkyl or halo.
Particularly R2 is hydrogen, methyl, fluoro or chloro.
More particularly R2 is C1-4alkyl or halo when R3 is hydrogen.
Preferably R3 is hydrogen, C1-6alkyl or halo.
More preferably R3 is hydrogen, C1-4alkyl or halo.
Particularly R3 is hydrogen, methyl, fluoro or chloro.
More particularly R3 is C1-4alkyl or halo when R2 is hydrogen.
Preferably R4 is hydrogen or C1-6alkoxy or R4 is aryl or heteroaryl optionally substituted by one or more groups selected from halo, cyano, C1-6alkyl, C1-6alkoxy, N,N-(C1-6alkyl)2amino or heterocyclyl.
More preferably R4 is hydrogen or C1-6alkoxy or R4 is aryl or heteroaryl optionally substituted by one or more groups selected from halo, cyano, C1-6alkyl, C1-6alkoxy, N,N-(C1-6alkyl)2amino, pyrrolidin-1-yl, piperidinyl, morpholino, piperazinyl, 4-C1-6alkylpiperazin-1-yl, homopiperazinyl-1-yl or 4-C1-6alkylhomopiperazinyl-1-yl.
More preferably R4 is hydrogen or C1-6alkoxy or R4 is aryl or heteroaryl optionally substituted by one or more groups selected from halo, cyano, C1-6alkyl, C1-6alkoxy, N,N-(C1-6alkyl)2amino, piperazinyl, morpholino or piperazinyl.
More preferably R4 is hydrogen or C1-4alkoxy or R4 is phenyl, thienyl, furyl, oxazolyl, isoxazolyl, pyrimidyl or pyridyl optionally substituted by one or two halo, cyano, C1-4alkyl, C1-4alkoxy, N,N-(C1-4alkyl)2amino, piperidinyl, morpholino or piperazinyl.
Particularly R4 is hydrogen or methoxy or R4 is phenyl, furyl, isoxazolyl or pyridyl optionally substituted by one or more groups selected from fluoro, chloro, cyano, methyl, methoxy, N,N-dimethylamino or morpholino.
More particularly R4 is hydrogen, methoxy, phenyl, 2-methylphenyl, 3-(N,N-dimethylamino)phenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-cyanophenyl, 3,4-dimethoxyphenyl, 3-morpholinophenyl, 2-furyl, 2-chloropyrid-5-yl, 2-morpholinopyrid-4-yl or isoxazol-5-yl.
Further more particularly R4 is pyridyl optionally substituted by a N,N-dimethylamino, N,N-diethylamino, pyrrolidin-1-yl, piperidino or morpholino group.
Even more particularly R4 is 2-morpholinopyrid-4-yl.
Preferably R5 is hydrogen.
Preferably G is N.
Preferably m is 2 or 3.
Particularly m is 1, 2 or 3.
Preferably q is 0 or 1.
When, as defined hereinbefore, any of the R1 or R4 groups defined hereinbefore which comprises a CH2 group which is attached to 2 carbon atoms or a CH3 group which is attached to a carbon atom may optionally bear on each said CH2 or CH3 group a substituent selected from hydroxy, amino, C1-6alkoxy, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino and heterocyclyl, suitable substituents so formed include, for example, substituted heterocyclylC1-6alkoxy groups such as 2-hydroxy-3-piperidinopropoxy and 2-hydroxy-3-morpholinopropoxy, substituted aminoC1-6alkoxy groups such as 3-amino-2-hydroxypropoxy, substituted N-C1-6alkylaminoC1-6alkoxy groups such as 2-hydroxy-3-methylaminopropoxy, substituted N,N-(C1-6alkyl)2aminoC1-6alkoxy groups such as 3-dimethylamino-2-hydroxypropoxy, 3-[N-(3-dimethylaminopropyl)-N-methylamino]propoxy and 3-[N-(3-dimethylaminopropyl)-N-methylamino]-2-hydroxypropoxy, substituted heterocyclylC1-6alkylamino groups such as 2-hydroxy-3-piperidinopropylamino and 2-hydroxy-3-morpholinopropylamino, substituted aminoC1-6alkylamino groups such as 3-amino-2-hydroxypropylamino, substituted N-C1-6alkylaminoC1-6alkylamino groups such as 2-hydroxy-3-methylaminopropylamino, substituted N,N-(C1-6alkyl)2aminoC1-6alkylamino groups such as 3-dimethylamino-2-hydroxypropylamino, 3-[N-(3-dimethylaminopropyl)-N-methylamino]propylamino and 3-[N-(3-dimethylaminopropyl)-N-methylamino]-2-hydroxypropylamino, substituted N-C1-6alkylaminoC1-6alkyl groups such as 2-dimethylaminoethylaminomethyl, 3-dimethylaminopropylaminomethyl, 3-dimethylamino-2,2-dimethylpropylaminomethyl, 2-morpholinoethylaminomethyl, 2-piperazin-1-ylethylaminomethyl and 3-morpholinopropylaminomethyl.
Preferred combinations of q and R4 are as follows.
When q is 0, preferably R4 is hydrogen or C1-6alkoxy or R4 is aryl or heteroaryl optionally substituted by one or more groups selected from halo, cyano, C1-6alkyl, C1-6alkoxy, N,N-(C1-6alkyl)2amino, piperazinyl, morpholino or piperazinyl. When q is 1, preferably R4 is hydrogen or C1-6alkoxy. When q is 2 preferably R4 is hydrogen.
When q is 0, more preferably R4 is hydrogen or C1-4alkoxy or R4 is phenyl, thienyl, furyl, oxazolyl, isoxazolyl, pyrimidyl or pyridyl optionally substituted by one or two halo, cyano, C1-4alkyl, C1-4alkoxy, N,N-(C1-4alkyl)2amino, piperazinyl, morpholino or piperazinyl groups. When q is 1, more preferably R4 is hydrogen or C1-4alkoxy.
When q is 0, particularly R4 is hydrogen or methoxy or R4 is phenyl, furyl, isoxazolyl or pyridyl optionally substituted by one or more groups selected from fluoro, chloro, cyano, methyl, methoxy, N,N-dimethylamino or morpholino. When q is 1, particularly R4 is hydrogen or methoxy.
When q is 0, more particularly R4 is hydrogen, methoxy, phenyl, 2-methylphenyl, 3-(N,N-dimethylamino)phenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-cyanophenyl, 3,4-dimethoxyphenyl, 3-morpholinophenyl, 2-furyl, 2-chloropyrid-5-yl, 2-morpholinopyridyl-4-yl or isoxazol-5-yl.
Preferred combinations of R1 and m are as follows.
When m is 2, preferably R1 is C1-6alkoxy, heterocyclylC1-6alkoxy or heteroarylC1-6alkoxy. When m is 3, preferably R1 is C1-6alkoxy.
When m is 2, more preferably R1 is C1-4alkoxy, morpholinylC2-4alkoxy, pyrrolidinylC2-4alkoxy or pyridylC2-4alkoxy. When m is 3, preferably R1 is C1-4alkoxy.
When m is 2 particularly R1 is methoxy, 2-morpholinoethoxy, 3-morpholinopropoxy, 2-pyrrolidin-1-ylethoxy or 3-pyrid-3-ylpropoxy. When m is 3, particularly R1 is methoxy.
When m is 2 more particularly (R1)m is 6,7-dimethoxy, 6-methoxy-7-[2-morpholinoethoxy], 6-methoxy-7-[3-morpholinopropoxy], 6-methoxy-7-(2-pyrrolidin-1-ylethoxy) or 6-methoxy-7-(3-pyrid-3-ylpropoxy). When m is 3, more particularly (R1)m is 6, 7, 8-trimethoxy.
In one embodiment of the invention R4 is hydrogen, hydroxy, C1-6alkyl, C1-6alkoxy, amino, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, hydroxyC2-6alkoxy, C1-6alkoxy-C2-6alkoxy, aminoC2-6alkoxy, N-C1-6alkylaminoC2-6alkoxy or N,N-(C1-6alkyl)2amino-C2-6alkoxy.
In a further embodiment of the invention R4 is of the Formula:
xe2x80x94Kxe2x80x94Jxe2x80x83xe2x80x83(IC)
wherein J is aryl, heteroaryl or heterocyclyl and K is a bond, oxy, imino, N-(C1-6alkyl)imino, oxyC1-6alkylene, iminoC1-6alkylene, N-(C1-6alkyl)iminoC1-6alkylene, xe2x80x94NHC(O)xe2x80x94, xe2x80x94SO2NHxe2x80x94, xe2x80x94NHSO2xe2x80x94 or xe2x80x94NHC(O)xe2x80x94C1-6alkylene-, and any aryl, heteroaryl or heterocyclyl may be optionally substituted by one or more groups selected from hydroxy, halo, trifluoromethyl, cyano, mercapto, nitro, amino, carboxy, carbamoyl, formyl, sulphamoyl, C1-6alkyl, C2-6alkenyl, C2-6alkynyl, C1-6alkoxy, xe2x80x94O-(C1-3alkyl)-Oxe2x80x94, C1-6alkylS(O)nxe2x80x94 (wherein n is 0-2), N-C1-6alkylamino, N,N-(C1-6alkyl)2amino, C1-6alkoxycarbonyl, N-C1-6alkylcarbamoyl, N,N-(C1-6alkyl)2carbamoyl, C2-6alkanoyl, C1-6alkanoyloxy, C1-6alkanoylamino, N-C1-6alkylsulphamoyl, N,N-(C1-6alkyl)2sulphamoyl, C1-6alkylsulphonylamino and C1-6alkylsulphonyl-N-(C1-6alkyl)amino, or any aryl, heteroaryl or heterocyclyl may be optionally substituted with one or more groups of the Formula (IAxe2x80x2) or (IBxe2x80x2) wherein A1, B1, D1 and E1 are as defined for Formula (I).
In another embodiment of the invention G is CH.
Another aspect of the present invention provides a compound of the Formula (I) (as depicted above) wherein:
R1 is hydroxy, halo, C1-6alkoxy, N,N-(C1-6alkyl)2aminoC1-6alkyl, N,N-(C1-6alkyl)2carbamoylC1-6alkoxy, N,N-(C1-6alkyl)2aminoC1-6alkoxy, C1-6alkylS(O)2xe2x80x94C1-6alkoxy, N,N-(C1-6alkyl)2amino-N-(C1-6alkyl)C1-6alkylamino, N,N-(C1-6alkyl)2aminoC1-6alkylaminoC1-6alkyl, heterocyclylC1-6alkyl, heterocyclyl-C1-6alkoxy, heterocyclyloxy, heterocyclylC1-6alkylaminoC1-6alkyl or heteroarylC1-6alkoxy;
R2 is hydrogen, C1-6alkyl or halo;
R3 is hydrogen, C1-6alkyl or halo;
R4 is hydrogen or C1-6alkoxy or R4 is aryl or heteroaryl optionally substituted by one or more groups selected from halo, cyano, C1-6alkyl, C1-6alkoxy, N,N-(C1-6alkyl)2amino or heterocyclyl;
R5 is hydrogen;
G is N;
m is 1, 2 or 3; and
q is 0 or 1;
or a pharmaceutically acceptable salt, or an in vivo cleavable ester thereof.
A further aspect of the present invention provides a compound of the Formula (I) (as depicted above) wherein:
R1 is hydroxy, halo, C1-6alkoxy, N,N-(C1-6alkyl)2aminoC1-6alkyl, N,N-(C1-6alkyl)2carbamoylC1-6alkoxy, N,N-(C1-6alkyl)2aminoC1-6alkoxy, C1-6alkylS(O)2xe2x80x94C1-6alkoxy, N,N-(C1-6alkyl)2amino-N-(C1-6alkyl)C1-6alkylamino, N,N-(C1-6alkyl)2aminoC1-6alkylaminoC1-6alkyl, piperazin-1-ylC1-6alkyl, 4-C1-6alkylpiperazin-1-ylC1-6alkyl, homopiperazinyl-1-ylC1-6alkyl, 4-C1-6alkylhomopiperazinyl-1-ylC1-4alkyl, pyrrolidinylC1-6alkoxy, piperidinylC1-6alkoxy, N-(C1-6alkyl)pyrrolidinylC1-6alkoxy, N-(C1-6alkyl)piperidinylC1-6alkoxy, morpholinylC1-6alkoxy, piperazinylC1-6alkoxy, N-(C1-6alkyl)piperazinylC1-6alkoxy, homopiperazinylC1-6alkoxy, N-(C1-6alkyl)homopiperazinylC1-6alkoxy, pyrrolidinyloxy, piperidinyloxy, morpholinylC1-6alkylaminoC1-6alkyl or pyridylC1-6alkoxy;
R2 is hydrogen, C1-6alkyl or halo;
R3 is hydrogen, C1-6alkyl or halo;
R4 is hydrogen or C1-6alkoxy or R4 is aryl or heteroaryl optionally substituted by one or more groups selected from halo, cyano, C1-6alkyl, C1-6alkoxy, N,N-(C1-6alkyl)2amino, pyrrolidin-1-yl, piperidinyl, morpholino, piperazinyl, 4-C1-6alkylpiperazin-1-yl, homopiperazinyl-1-yl or 4-C1-6alkylhomopiperazinyl-1-yl;
R5 is hydrogen;
G is N;
m is 1, 2 or 3; and
q is 0 or 1;
or a pharmaceutically acceptable salt, or an in vivo cleavable ester thereof.
A further aspect of the present invention provides a compound of the Formula (I) (as depicted above) wherein:
R1 is C1-6alkoxy, heterocyclylC1-6alkoxy or heteroarylC1-6alkoxy;
R2 is hydrogen, C1-6alkyl or halo;
R3 is hydrogen, C1-6alkyl or halo;
R4 is hydrogen or C1-6alkoxy or R4 is aryl or heteroaryl optionally substituted by one or more groups selected from halo, cyano, C1-6alkyl, C1-6alkoxy or N,N-(C1-6alkyl)2amino;
R5 is hydrogen;
G is N;
m is 2 or 3; and
q is 0 or 1;
or a pharmaceutically acceptable salt, or an in vivo cleavable ester thereof.
A further aspect of the present invention provides a compound of the Formula (I) (as depicted above) wherein:
R1 is C1-6alkoxy, morpholinylC1-6alkoxy, pyrrolidinylC1-6alkoxy or pyridylC1-6alkoxy;
R2 is hydrogen, C1-6alkyl or halo;
R3 is hydrogen, C1-6alkyl or halo;
R4 is hydrogen or C1-6alkoxy or R4 is aryl or heteroaryl optionally substituted by one or more groups selected from halo, cyano, C1-6alkyl, C1-6alkoxy, N,N-(C1-6alkyl)2amino, piperidinyl, morpholino or piperazinyl;
R5 is hydrogen;
G is N;
m is 2 or 3; and
q is 0 or 1;
or a pharmaceutically acceptable salt, or an in vivo cleavable ester thereof.
An additional aspect of the present invention provides a compound of the Formula (I) (as depicted above) wherein:
R1 is C1-4alkoxy, morpholinylC2-4alkoxy, pyrrolidinylC2-4alkoxy or pyridylC2-4alkoxy;
R2 is hydrogen, methyl, fluoro or chloro;
R3 is hydrogen, methyl, fluoro or chloro;
R4 is hydrogen or methoxy or R4 is phenyl, furyl, isoxazolyl or pyridyl optionally substituted by one or more groups selected from fluoro, chloro, cyano, methyl, methoxy, N,N-dimethylamino or morpholino;
R5 is hydrogen;
G is N;
m is 2 or 3; and
q is 0 or 1;
or a pharmaceutically acceptable salt, or an in vivo cleavable ester thereof
A further additional aspect of the present invention provides a compound of the Formula (I) (as depicted above) wherein:
R1 is methoxy, 2-morpholinoethoxy, 3-morpholinopropoxy, 2-pyrrolidin-1-ylethoxy or 3-pyrid-3-ylpropoxy;
R2 is hydrogen, methyl, fluoro or chloro;
R3 is hydrogen, methyl, fluoro or chloro;
R4 is hydrogen, methoxy, phenyl, 2-methylphenyl, 3-(N,N-dimethylamino)phenyl, 3-fluorophenyl, 3-methoxyphenyl, 4-cyanophenyl, 3,4-dimethoxyphenyl, 3-morpholinophenyl, 2-furyl, 2-chloropyrid-5-yl, 2-morpholinopyrid-4-yl or isoxazol-5-yl;
R5 is hydrogen;
G is N;
m is 2 or 3; and
q is 0 or 1;
or a pharmaceutically acceptable salt, or an in vivo cleavable ester thereof.
A further additional aspect of the present invention provides a compound of the Formula (I) (as depicted above) wherein:
R1 is methoxy, 2-dimethylaminoethoxy, 2-diethylaminoethoxy, 2-diisopropylaminoethoxy, 3-dimethylaminopropoxy, 3-diethylaminopropoxy, 2-morpholinoethoxy, 3-morpholinopropoxy, 2-piperidinoethoxy, N-methylpiperidin-2-ylmethoxy, N-methylpiperidin-3-ylmethoxy, 2-pyrrolidin-1-ylethoxy, 2-(N-methylpyrrolidin-2-yl)ethoxy, N-methyl-5-oxopyrrolidin-2-ylmethoxy, 3-pyrrolidin-1-ylpropoxy, 2-(2-oxoimidazolidin-1-yl)ethoxy, 2-(4-methylpiperazin-1-yl)ethoxy or 3-pyrid-3-ylpropoxy;
R2 is hydrogen, methyl, fluoro or chloro;
R3 is hydrogen, methyl, fluoro or chloro;
R4 is pyridyl optionally substituted by a N,N-dimethylamino, N,N-diethylamino, pyrrolidin-1-yl, piperidino or morpholino group.
R5 is hydrogen;
G is N;
m is 1, 2 or 3; and
q is 0;
or a pharmaceutically acceptable salt, or an in vivo cleavable ester thereof.
Preferred compounds are those of Examples 1-86 or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof.
Especially preferred compounds are those of Examples 18, 20, 23, 26, 31, 33, 34, 36, 40, 44, 45 or 48 or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof.
More especially preferred compounds are those of Examples 56, 58, 60, 61, 62, 63, 64, 65, 66, 68, 69 or 73 or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof.
A further especially preferred compound of the invention is, for example:xe2x80x94
4-(3-benzamido-4-fluoroanilino)-6,7-dimethoxyquinazoline,
6-(2-diisopropylaminoethoxy)-7-methoxy-4-[2-methyl-5-(2-morpholinopyridine-4-carboxamido)anilino]quinazoline,
6-(2-dimethylaminoethoxy)-7-methoxy-4-[2-methyl-5-(2-morpholinopyridine-4-carboxamido)anilino]quinazoline or
6-(3-pyrrolidin-1-ylpropoxy)-7-methoxy-4-[2-methyl-5-(2-morpholinopyridine-4-carboxamido)anilino]quinazoline;
or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof.
A further especially preferred compound of the invention is, for example:xe2x80x94
4-(3-benzamido-4-fluoroanilino)-6,7-dimethoxyquinoline;
or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof.
A suitable pharmaceutically acceptable salt of a compound of the Formula (I) is, for example, an acid-addition salt of a compound of the Formula (I) which is sufficiently basic, for example an acid-addition salt with an inorganic or organic acid such as hydrochloric, hydrobromic, sulphuric, trifluoroacetic, citric or maleic acid; or, for example a salt of a compound of the Formula (I) which is sufficiently acidic, for example an alkali or alkaline earth metal salt such as a calcium or magnesium salt, or an ammonium salt, or a salt with an organic base such as methylamine, dimethylamine, trimethylamine, piperidine, morpholine or tris-(2-hydroxyethyl)amine.
Various forms of prodrugs are known in the art. For examples of such prodrug derivatives, see:
a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) and Methods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al. (Academic Press, 1985);
b) A Textbook of Drug Design and Development, edited by Krogsgaard-Larsen and H. Bundgaard, Chapter 5 xe2x80x9cDesign and Application of Prodrugsxe2x80x9d, by H. Bundgaard p. 113-191 (1991);
c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);
d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285 (1988); and
e) N. Kakeya, et al., Chem. Pharm. Bull., 32, 692 (1984).
Examples of such pro-drugs may be used to form in vivo cleavable esters of a compound of the Formula (I). An in vivo cleavable ester of a compound of the Formula (I) containing a carboxy group is, for example, a pharmaceutically-acceptable ester which is cleaved in the human or animal body to produce the parent acid. Suitable pharmaceutically-acceptable esters for carboxy include C1-6alkoxymethyl esters, for example methoxymethyl; C1-6alkanoyloxymethylesters, for example pivaloyloxymethyl; phthalidyl esters; C3-8cycloalkoxycarbonyloxyC1-6alkyl esters, for example 1-cyclohexylcarbonyloxyethyl; 1,3-dioxolan-2-ylmethyl esters, for example 5-methyl-1,3-dioxolan-2-ylmethyl; and C1-6alkoxycarbonyloxyethyl esters, for example 1-methoxycarbonyloxyethyl; and may be formed at any carboxy group in the compounds of this invention.
In order to use a compound of the Formula (I), or a pharmaceutically acceptable salt or in vivo cleavable ester thereof, for the therapeutic treatment (including prophylactic treatment) of mammals including humans, it is normally formulated in accordance with standard pharmaceutical practice as a pharmaceutical composition.
According to this aspect of the invention there is provided a pharmaceutical composition which comprises an amide derivative of the Formula (I), or a pharmaceutically acceptable salt or in vivo cleavable ester thereof, as defined hereinbefore in association with a pharmaceutically acceptable diluent or carrier.
The compositions of the invention may be in a form suitable for oral use (for example as tablets, lozenges, hard or soft capsules, aqueous or oily suspensions, emulsions, dispersible powders or granules, syrups or elixirs), for topical use (for example as creams, ointments, gels, or aqueous or oily solutions or suspensions), for administration by inhalation (for example as a finely divided powder or a liquid aerosol), for administration by insufflation (for example as a finely divided powder) or for parenteral administration (for example as a sterile aqueous or oily solution for intravenous, subcutaneous, intramuscular or intramuscular dosing or as a suppository for rectal dosing).
The compositions of the invention may be obtained by conventional procedures using conventional pharmaceutical excipients, well known in the art. Thus, compositions intended for oral use may contain, for example, one or more colouring, sweetening, flavouring and/or preservative agents.
Suitable pharmaceutically acceptable excipients for a tablet formulation include, for example, inert diluents such as lactose, sodium carbonate, calcium phosphate or calcium carbonate, granulating and disintegrating agents such as corn starch or algenic acid; binding agents such as starch; lubricating agents such as magnesium stearate, stearic acid or talc; preservative agents such as ethyl or propyl p-hydroxybenzoate, and anti-oxidants, such as ascorbic acid. Tablet formulations may be uncoated or coated either to modify their disintegration and the subsequent absorption of the active ingredient within the gastrointestinal tract, or to improve their stability and/or appearance, in either case, using conventional coating agents and procedures well known in the art.
Compositions for oral use may be in the form of hard gelatin capsules in which the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with water or an oil such as peanut oil, liquid paraffin, or olive oil.
Aqueous suspensions generally contain the active ingredient in finely powdered form together with one or more suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinyl-pyrrolidone, gum tragacanth and gum acacia: dispersing or wetting agents such as lecithin or condensation products of an alkylene oxide with fatty acids (for example polyoxethylene stearate), or condensation products of ethylene oxide with long chain aliphatic alcohols, for example heptadecaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives (such as ethyl or propyl p-hydroxybenzoate, anti-oxidants (such as ascorbic acid), colouring agents, flavouring agents, and/or sweetening agents (such as sucrose, saccharine or aspartame).
Oily suspensions may be formulated by suspending the active ingredient in a vegetable oil (such as arachis oil, olive oil, sesame oil or coconut oil) or in a mineral oil (such as liquid paraffin). The oily suspensions may also contain a thickening agent such as beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set out above, and flavouring agents may be added to provide a palatable oral preparation. These compositions may be preserved by the addition of an anti-oxidant such as ascorbic acid.
Dispersible powders and granules suitable for preparation of an aqueous suspension by the addition of water generally contain the active ingredient together with a dispersing or wetting agent, suspending agent and one or more preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients such as sweetening, flavouring and colouring agents, may also be present.
The pharmaceutical compositions of the invention may also be in the form of oil-in-water emulsions. The oily phase may be a vegetable oil, such as olive oil or arachis oil, or a mineral oil, such as for example liquid paraffin or a mixture of any of these. Suitable emulsifying agents may be, for example, naturally-occurring gums such as gum acacia or gum tragacanth, naturally-occurring phosphatides such as soya bean, lecithin, an esters or partial esters derived from fatty acids and hexitol anhydrides (for example sorbitan monooleate) and condensation products of the said partial esters with ethylene oxide such as polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening, flavouring and preservative agents.
Syrups and elixirs may be formulated with sweetening agents such as glycerol, propylene glycol, sorbitol, aspartame or sucrose, and may also contain a demulcent, preservative, flavouring and/or colouring agent.
The pharmaceutical compositions may also be in the form of a sterile injectable aqueous or oily suspension, which may be formulated according to known procedures using one or more of the appropriate dispersing or wetting agents and suspending agents, which have been mentioned above. A sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic parentally-acceptable diluent or solvent, for example a solution in 1,3-butanediol.
Suppository formulations may be prepared by mixing the active ingredient with a suitable non-irritating excipient which is solid at ordinary temperatures but liquid at the rectal temperature and will therefore melt in the rectum to release the drug. Suitable excipients include, for example, cocoa butter and polyethylene glycols.
Topical formulations, such as creams, ointments, gels and aqueous or oily solutions or suspensions, may generally be obtained by formulating an active ingredient with a conventional, topically acceptable, vehicle or diluent using conventional procedures well known in the art.
Compositions for administration by insufflation may be in the form of a finely divided powder containing particles of average diameter of, for example, 30 xcexcm or much less, the powder itself comprising either active ingredient alone or diluted with one or more physiologically acceptable carriers such as lactose. The powder for insufflation is then conveniently retained in a capsule containing, for example, 1 to 50 mg of active ingredient for use with a turbo-inhaler device, such as is used for insufflation of the known agent sodium cromoglycate.
Compositions for administration by inhalation may be in the form of a conventional pressurised aerosol arranged to dispense the active ingredient either as an aerosol containing finely divided solid or liquid droplets. Conventional aerosol propellants such as volatile fluorinated hydrocarbons or hydrocarbons may be used and the aerosol device is conveniently arranged to dispense a metered quantity of active ingredient.
For further information on Formulation the reader is referred to Chapter 25.2 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.
The amount of active ingredient that is combined with one or more excipients to produce a single dosage form will necessarily vary depending upon the host treated and the particular route of administration. For example, a formulation intended for oral administration to humans will generally contain, for example, from 0.5 mg to 2 g of active agent compounded with an appropriate and convenient amount of excipients which may vary from about 5 to about 98 percent by weight of the total composition. Dosage unit forms will generally contain about 1 mg to about 500 mg of an active ingredient. For further information on Routes of Administration and Dosage Regimes the reader is referred to Chapter 25.3 in Volume 5 of Comprehensive Medicinal Chemistry (Corwin Hansch; Chairman of Editorial Board), Pergamon Press 1990.
The size of the dose for therapeutic or prophylactic purposes of a compound of the Formula (I) will naturally vary according to the nature and severity of the conditions, the age and sex of the animal or patient and the route of administration, according to well known principles of medicine.
In using a compound of the Formula (I) for therapeutic or prophylactic purposes it will generally be administered so that a daily dose in the range, for example, 0.5 mg to 75 mg per kg body weight, preferably 0.5 mg to 40 mg per kg body weight, is received, given if required in divided doses. In general lower doses will be administered when a parenteral route is employed. Thus, for example, for intravenous administration, a dose in the range, for example, 0.5 mg to 30 mg per kg body weight will generally be used. Similarly, for administration by inhalation, a dose in the range, for example, 0.5 mg to 25 mg per kg body weight will be used. Oral administration is however preferred, particularly in tablet form. Typically, unit dosage forms will contain about 1 mg to 500 mg of a compound of this invention.
The compounds of this invention may be used in combination with other drugs and therapies used in the treatment of disease states which would benefit from the inhibition of cytokines, in particular TNF and IL-1. For example, the compounds of the Formula (I) could be used in combination with drugs and therapies used in the treatment of rheumatoid arthritis, asthma, irritable bowel disease, multiple sclerosis, AIDS, septic shock, congestive heart failure, ischaemic heart disease, psoriasis and the other disease states mentioned earlier in this specification.
For example, by virtue of their ability to inhibit cytokines, the compounds of the Formula (I) are of value in the treatment of certain inflammatory and non-inflammatory diseases which are currently treated with a cyclooxygenase-inhibitory non-steroidal anti-inflammatory drug (NSAID) such as indomethacin, ketorolac, acetylsalicylic acid, ibuprofen, sulindac, tolmetin and piroxicam. Co-administration of a compound of the Formula (I) with a NSAID can result in a reduction of the quantity of the latter agent needed to produce a therapeutic effect. Thereby the likelihood of adverse side-effects from the NSAID such as gastrointestinal effects are reduced. Thus according to a further feature of the invention there is provided a pharmaceutical composition which comprises a compound of the Formula (I), or a pharmaceutically acceptable salt or in vivo cleavable ester thereof, in conjunction or admixture with a cyclooxygenase inhibitory non-steroidal anti-inflammatory agent, and a pharmaceutically acceptable diluent or carrier.
The compounds of the invention may also be used with anti-inflammatory agents such as an inhibitor of the enzyme 5-lipoxygenase (such as those disclosed in European Patent Applications Nos. 0351194, 0375368, 0375404, 0375452, 0375457, 0381375, 0385662, 0385663, 0385679, 0385680).
The compounds of the Formula (I) may also be used in the treatment of conditions such as rheumatoid arthritis in combination with antiarthritic agents such as gold, methotrexate, steroids and penicillinamine, and in conditions such as osteoarthritis in combination with steroids.
The compounds of the present invention may also be administered in degradative diseases, for example osteoarthritis, with chondroprotective, anti-degradative and/or reparative agents such as Diacerhein, hyaluronic acid formulations such as Hyalan, Rumalon, Arteparon and glucosamine salts such as Antril.
The compounds of the Formula (I) may be used in the treatment of asthma in combination with antiasthmatic agents such as bronchodilators and leukotriene antagonists.
If formulated as a fixed dose such combination products employ the compounds of this invention within the dosage range described herein and the other pharmaceutically-active agent within its approved dosage range. Sequential use is contemplated when a combination formulation is inappropriate.
Although the compounds of the Formula (I) are primarily of value as therapeutic agents for use in warm-blooded animals (including man), they are also useful whenever it is required to inhibit the effects of cytokines. Thus, they are useful as pharmacological standards for use in the development of new biological tests and in the search for new pharmacological agents.
According to a further aspect of the present invention, there is provided a process for preparing a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof, which process (wherein R1, R2, R3, R4, R5, G, m and q are as defined for Formula (I) unless otherwise stated) comprises of:
a) reacting an aniline of the Formula (II): 
with an acyl compound of the Formula (III): 
wherein L is a displaceable group as defined below;
b) reacting an activated heteroaryl of the Formula (IV): 
xe2x80x83wherein L is a displaceable group as defined below, with an aniline of the Formula (V): 
xe2x80x83or c) for the preparation of a compound of the Formula (I) wherein R1 or a substituent on R4 is C1-6alkoxy or substituted C1-6alkoxy, C1-6alkylS-, N-C1-6alkylamino, N,N-(C1-6alkyl)2amino or substituted C1-6alkylamino, the alkylation, conveniently in the presence of a suitable base as defined below, of an amide derivative of the Formula (I) wherein R1 or a substituent on R4 is hydroxy, mercapto or amino as appropriate; and thereafter if necessary:
i) converting a compound of the Formula (I) into another compound of the Formula (I);
ii) removing any protecting groups; and
iii) forming a pharmaceutically acceptable salt or in vivo cleavable ester.
Specific reaction conditions for the above process variants are as follows:xe2x80x94
For process variant a) A suitable displaceable group L is, for example, a halogeno, activated phenoxy group or sulphonyloxy group, for example a chloro, bromo, pentafluorophenoxy or methanesulphonyloxy or toluene-4-sulphonyloxy group. Especially preferred displaceable groups are chloro and pentafluorophenoxy.
Anilines of the Formula (II) and acyl compounds of the Formula (III) may be reacted together in a suitable inert solvent or diluent, for example dichloromethane, acetonitrile, butanol, tetramethylene sulphone, tetrahydrofuran 1,2-dimethoxyethane, N,N-dimethylformamide, N,N-dimethylacetamide or N-methylpyrrolidin-2-one, optionally in the presence of a suitable base, and at a temperature in the range, for example, 0xc2x0 to 50xc2x0 C., conveniently at or near room temperature.
A suitable base is, for example, an alkali or alkaline earth metal carbonate, alkoxide, hydroxide or hydride, for example sodium carbonate, potassium carbonate, sodium ethoxide, potassium butoxide, sodium hydroxide, potassium hydroxide, sodium hydride or potassium hydride, or an organometallic base such as an alkyl-lithium, for example n-butyl-lithium, or a dialkylamino-lithium, for example lithium di-isopropylamide, or, for example, an organic amine base such as, for example, pyridine, 2,6-lutidine, collidine, 4-dimethylaminopyridine, triethylamine, morpholine or diazabicyclo[5.4.0]undec-7-ene.
Anilines of the Formula (II) may be prepared by the reaction of the activated heteroaryl of the Formula (IV) according to the following scheme: 
wherein Q is xe2x80x94NH2 or, if R2 and R3 are not identical and a regiospecific reaction is desired, Q can be amino protected by a suitable protecting group (such as those defined below) or nitro, whereafter the protecting group is removed, or the nitro group is reduced (for example with iron powder and acetic acid) to generate the aniline of the Formula (II).
Activated heteroaryls of the Formula (IV) are known compounds, are commercially available or are prepared by processes known in the art. For example where L is chloro as in Formula (IVB) or pentafluorophenoxy as in Formula (IVC), suitable compounds of the Formula (IV) may be prepared by the following scheme from compounds of the Formula (IVA) which are known compounds, are commercially available or are prepared by processes known in the art.: 
For process variant b) A suitable displaceable group L is as defined above.
Activated heteroaryls of the formula (IV) and anilines of the Formula (V) may be reacted together in the presence of a protic solvent, for example, isopropanol, in the presence of an acid, for example hydrogen chloride gas in diethyl ether, or hydrochloric acid, and at a temperature in the range, for example, 0xc2x0 to 150xc2x0 C., conveniently at or near reflux.
Anilines of the Formula (V) are, known compounds, are commercially available, or are made by processes known in the art. For example, anilines of the Formula (V) may be prepared according to the following scheme: 
wherein Q is as defined above.
Compounds of the Formulae (IIB), (III), (VA) and (VB) are known compounds, are commercially available or are prepared by processes known in the art.
For process variant c) A suitable alkylating agent is, for example, any agent known in the art for the alkylation of hydroxy to alkoxy or substituted alkoxy, or for the alkylation of mercapto to alkylthio, or for the alkylation of amino to alkylamino or substituted alkylamino, for example an alkyl or substituted alkyl halide, for example a C1-6alkyl chloride, bromide or iodide or a substituted C1-6alkyl chloride, bromide or iodide, in the presence of a suitable base and in a suitable inert solvent or diluent as defined above for process variant a).
The reaction is conveniently carried out at a temperature in the range, for example, 10 to 150xc2x0 C., preferably in the range 20 to 80xc2x0 C.
Any necessary protecting groups may in general be chosen from any of the groups described in the literature or known to the skilled chemist as appropriate for the protection of the group in question and may be introduced by conventional methods Protecting groups may be removed by any convenient method as described in the literature or known to the skilled chemist as appropriate for the removal of the protecting group in question, such methods being chosen so as to effect removal of the protecting group with minimum disturbance of groups elsewhere in the molecule.
Specific examples of protecting groups are given below for the sake of convenience, in which xe2x80x9clowerxe2x80x9d, as in, for example, lower alkyl, signifies that the group to which it is applied preferably has 1-4 carbon atoms. It will be understood that these examples are not exhaustive. Where specific examples of methods for the removal of protecting groups are given below these are similarly not exhaustive. The use of protecting groups and methods of deprotection not specifically mentioned would be available to the skilled chemist and are within the scope of the invention.
A carboxy protecting group may be the residue of an ester-forming aliphatic or arylaliphatic alcohol or of an ester-forming silanol (the said alcohol or silanol preferably containing 1-20 carbon atoms). Examples of carboxy protecting groups include esters involving straight or branched chain C1-12alkyl groups (for example isopropyl, tert-butyl); lower alkoxy lower alkyl groups (for example methoxymethyl, ethoxymethyl, isobutoxymethyl); lower aliphatic acyloxy lower alkyl groups, (for example acetoxymethyl, propionyloxymethyl, butyryloxymethyl, pivaloyloxymethyl); lower alkoxycarbonyloxy lower alkyl groups (for example 1-methoxycarbonyloxyethyl, 1-ethoxycarbonyloxyethyl); aryl lower alkyl groups (for example benzyl, p-methoxybenzyl, o-nitrobenzyl, p-nitrobenzyl, benzhydryl and phthalidyl); tri(lower alkyl)silyl groups (for example trimethylsilyl and tert-butyldimethylsilyl); tri(lower alkyl)silyl lower alkyl groups (for example trimethylsilylethyl); and C2-6alkenyl groups (for example allyl and vinylethyl).
Methods particularly appropriate for the removal of carboxyl protecting groups include for example acid-, base-, metal- or enzymically-catalysed hydrolysis.
Examples of hydroxy protecting groups include ethers involving lower alkyl groups (for example tert-butyl), lower alkenyl groups (for example allyl); lower alkanoyl groups (for example acetyl); lower alkoxycarbonyl groups (for example tert-butoxycarbonyl); lower alkenyloxycarbonyl groups (for example allyloxycarbonyl); aryl lower alkoxycarbonyl groups (for example benzoyloxycarbonyl, p-methoxybenzyloxycarbonyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl); tri lower alkylsilyl groups (for example trimethylsilyl, tert-butyldimethylsilyl) and aryl lower alkyl groups (for example benzyl).
Examples of amino protecting groups include amides or amines involving formyl, aralkyl groups (for example benzyl and substituted benzyl, p-methoxybenzyl, nitrobenzyl and 2,4-dimethoxybenzyl, and triphenylmethyl); di-p-anisylmethyl and furylmethyl groups; lower alkoxycarbonyl (for example tert-butoxycarbonyl); lower alkenyloxycarbonyl (for example allyloxycarbonyl); aryl lower alkoxycarbonyl groups (for example benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl; trialkylsilyl (for example trimethylsilyl and tert-butyldimethylsilyl); alkylidene (for example methylidene); benzylidene and substituted benzylidene groups.
Methods appropriate for removal of hydroxy and amino protecting groups include, for example, acid-, base-, metal- or enzymically-catalysed hydrolysis for groups such as p-nitrobenzyloxycarbonyl, hydrogenation for groups such as benzyl and photolytically for groups such as o-nitrobenzyloxycarbonyl.
The reader is referred to Advanced Organic Chemistry, 4th Edition, by Jerry March, published by John Wiley and Sons 1992, for general guidance on reaction conditions and reagents. The reader is referred to Protective Groups in Organic Synthesis, 2nd Edition, by Green et al., published by John Wiley and Sons for general guidance on protecting groups.
According to a further aspect of the present invention there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, for use in a method of treatment of the human or animal body by therapy.
In a further aspect of the present invention there is provided a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, for use as a medicament.
In a further aspect the present invention provides the use of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, in the manufacture of a medicament for use in the treatment of diseases or medical conditions mediated by cytokines.
In a further aspect the present invention provides a method of treating diseases or medical conditions mediated by cytokines which comprises administering to a warm-blooded animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore.
In a further aspect the present invention provides the use of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, in the manufacture of a medicament for use in the treatment of diseases or medical conditions mediated by TNF, IL-1, IL-6 or IL-8.
In a further aspect the present invention provides a method of treating diseases or medical conditions mediated by TNF, IL-1, IL-6 or IL-8 which comprises administering to a warm-blooded animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore.
In a further aspect the present invention provides the use of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, in the manufacture of a medicament for use in the treatment of diseases or medical conditions mediated by TNF.
In a further aspect the present invention provides a method of treating diseases or medical conditions mediated by TNF which comprises administering to a warm-blooded animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore.
In a further aspect the present invention provides the use of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, in the manufacture of a medicament for use in inhibiting TNF, IL-1, IL-6 or IL-8.
In a further aspect the present invention provides a method of inhibiting TNF, IL-1, IL-6 or IL-8 which comprises administering to a warm-blooded animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore.
In a further aspect the present invention provides the use of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, in the manufacture of a medicament for use in inhibiting TNF.
In a further aspect the present invention provides a method of inhibiting TNF which comprises administering to a warm-blooded animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore.
In a further aspect the present invention provides the use of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, in the manufacture of a medicament for use in the treatment of diseases or medical conditions mediated by p38 kinase.
In a further aspect the present invention provides a method of treating diseases or medical conditions mediated by p38 kinase which comprises administering to a warm-blooded animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore.
In a further aspect the present invention provides the use of a compound of the. Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, in the manufacture of a medicament for use in the production of a p38 kinase inhibitory effect.
In a further aspect the present invention provides a method of providing a p38 kinase inhibitory effect which comprises administering to a warm-blooded animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore.
In a further aspect the present invention provides the use of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore, in the manufacture of a medicament for use in the treatment of rheumatoid arthritis, asthma, irritable bowel disease, multiple sclerosis, AIDS, septic shock, congestive heart failure, ischaemic heart disease or psoriasis.
In a further aspect the present invention provides a method of treating rheumatoid arthritis, asthma, irritable bowel disease, multiple sclerosis. AIDS, septic shock, congestive heart failure, ischaemic heart disease or psoriasis which comprises administering to a warm-blooded animal an effective amount of a compound of the Formula (I), or a pharmaceutically acceptable salt or an in vivo cleavable ester thereof as defined hereinbefore.
The following biological assays and Examples serve to illustrate the present invention.
Biological Assays
The following assays can be used to measure the p38 kinase-inhibitory, the TNF-inhibitory and anti-arthritic effects of the compounds of the present invention:
In Vitro Enzyme Assay
The ability of compounds of the invention to inhibit the enzyme p38 kinase was assessed. Activity of particular test compounds against each of the p38xcex1 and p38xcex2 isoforms of the enzyme was determined.
Human recombinant MKK6 (GenBank Accesion Number G1209672) was isolated from Image clone 45578 (Genomics, 1996, 33, 151) and utilised to produce protein in the form of a GST fusion protein in a pGEX vector using analogous procedures to those disclosed by J. Han et al., Journal of Biological Chemistry, 1996, 271 2886-2891. p38xcex1 (GenBank Accession Number G529039) and p38xcex2 (GenBank Accession Number G1469305) were isolated by PCR amplification of human lymphoblastoid cDNA (GenBank Accession Number GM1416) and human foetal brain cDNA [synthesised from mRNA (Clontech, catalogue no. 6525-1) using a Gibco superscript cDNA synthesis kit] respectively using oligonucleotides designed for the 5xe2x80x2 and 3xe2x80x2 ends of the human p38xcex1 and p38xcex2 genes using analogous procedures to those described by J. Han et al., Biochimica et Biophysica Acta, 1995, 1265, 224-227 and Y. Jiang et al., Journal of Biological Chemistry, 1996, 271, 17920-17926.
Both p38 protein isoforms were expressed in e coli in PET vectors. Human recombinant p38xcex1 and p38xcex2 isoforms were produced as 5xe2x80x2 c-myc, 6His tagged proteins. Both MKK6 and the p38 proteins were purified using standard protocols: the GST MKK6 was purified using a glutathione sepharose column and the p38 proteins were purified using nickel chelate columns.
The p38 enzymes were activated prior to use by incubation with MKK6 for 3 hours at 30xc2x0 C. The unactivated coli-expressed MKK6 retained sufficient activity to fully activate both isoforms of p38. The activation incubate comprised p38xcex1 (10 xcexcl of 10 mg/ml) or p38xcex2 (10 xcexcl of 5 mg/ml) together with MKK6 (10 xcexcl of 1 mg/ml), xe2x80x98Kinase bufferxe2x80x99 [100 xcexcl; pH 7.4 buffer comprising Tris (50 mM), EGTA (0.1 mM), sodium orthovanadate (0.1 mM) and xcex2-mercaptoethanol (0.1%)] and MgATP (30 xcexcl of 50 mM Mg(OCOCH3)2 and 0.5 mM ATP). This produced enough activated p38 enzyme for 3 Microtiter plates.
Test compounds were solubilised in DMSO and 10 xcexcl of a 1:10 diluted sample in xe2x80x98Kinase Bufferxe2x80x99 was added to a well in a Microtiter plate. For single dose testing, the compounds were tested at 10 xcexcM. xe2x80x98Kinase Assay Mixxe2x80x99 [30 xcexcl; comprising Myelin Basic Protein (Gibco BRL cat. no. 1322B-010; 1 ml of a 3.33 mg/ml solution in water), activated p38 enzyme (50 xcexcl) and xe2x80x98Kinase Bufferxe2x80x99 (2 ml)] was then added followed by xe2x80x98Labelled ATPxe2x80x99 [10 xcexcl; comprising 50 xcexcM ATP, 0.1 xcexcCi 33P ATP (Amersham International cat. no. BF1000) and 50 mM Mg(OCOCH3)2]. The plates were incubated at room temperature with gentle agitation. Plates containing p38xcex1 were incubated for 90 min and plates containing p38xcex2 were incubated for 45 min. Incubation was stopped by the addition of 50 xcexcl of 20% trichloroacetic acid (TCA). The precipitated protein was phosphorylated by p38 kinase and test compounds were assessed for their ability to inhibit this phosphorylation. The plates were filtered using a Canberra Packard Unifilter and washed with 2% TCA, dried overnight and counted on a Top Count scintillation counter.
Test compounds were tested initially at a single dose and active compounds were retested to allow IC50 values to be determined.
In Vitro Cell-Based Assays
(i) PBMC
The ability of compounds of this invention to inhibit TNFxcex1 production was assessed by using human peripheral blood mononuclear cells which synthesise and secrete TNFxcex1 when stimulated with lipopolysaccharide.
Peripheral blood mononuclear cells (PBMC) were isolated from heparinised (10 units/ml heparin) human blood by density centrifugation (Lymphoprep(trademark); Nycomed). Mononuclear cells were resuspended in culture medium [RPMI 1640 medium (Gibco) supplemented with 50 units/ml penicillin 50 xcexcg/ml streptomycin, 2 mM glutamine and 1% heat-inactivated human AB serum (Sigma H-1513)]. Compounds were solubilised in DMSO at a concentration of 50 mM, diluted 1:100 in culture medium and subsequently serial dilutions were made in culture medium containing 1% DMSO. PBMCs (2.4xc3x97105 cells in 160 xcexcl culture medium) were incubated with 20 xcexcl of varying concentrations of test compound (triplicate cultures) or 20 xcexcl culture medium containing 1% DMSO (control wells) for 30 minutes at 37xc2x0 C. in a humidified (5%CO2/95% air) incubator (Falcon 3072; 96 well flat-bottom tissue culture plates). 20 xcexcl lipopolysaccharide [LPS E. Coli 0111:B4 (Sigma L-4130), final concentration 10 xcexcg/ml] solubilised in culture medium was added to appropriate wells. 20 xcexcl culture medium was added to xe2x80x9cmedium alonexe2x80x9d control wells. Six xe2x80x9cLPS alonexe2x80x9d and four xe2x80x9cmedium alonexe2x80x9d controls were included on each 96 well plate. Varying concentrations of a known TNFxcex1 inhibitor were included in each test, i.e. an inhibitor of the PDE Type IV enzyme (for example see Semmler, J. Wachtel. H and Endres, S., Int. J. Immunopharmac. (1993), 15(3), 409-413) or an inhibitor of proTNFxcex1 convertase (for example, see McGeehan, G. M. et al. Nature (1994) 370, 558-561). Plates were incubated for 7 hours at 37xc2x0 C. (humidified incubator) after which 100 xcexcl of the supernatant was removed from each well and stored at xe2x88x9270xc2x0 C. (96 well round-bottom plates; Coming 25850). TNFxcex1 levels were determined in each sample using a human TNFxcex1 ELISA (see WO92/10190 and Current Protocols in Molecular Biology, vol 2 by Frederick M. Ausbel et al., John Wiley and Sons Inc.).             %      ⁢              xe2x80x83            ⁢      inhibition        =                                        xe2x80x83                    ⁢                                    (                              LPS                ⁢                                  xe2x80x83                                ⁢                alone                ⁢                                  -                                ⁢                medium                ⁢                                  xe2x80x83                                ⁢                alone                            )                        -                          (                              test                ⁢                                  xe2x80x83                                ⁢                concentration                ⁢                                  -                                ⁢                medium                ⁢                                  xe2x80x83                                ⁢                alone                            )                                                (                      LPS            ⁢                          xe2x80x83                        ⁢            alone            ⁢                          -                        ⁢            medium            ⁢                          xe2x80x83                        ⁢            alone                    )                    xc3x97      100        ⁢      
    ⁢      xe2x80x83  
(ii) Human Whole Blood
The ability of the compounds of this invention to inhibit TNFxcex1 production was also assessed in a human whole blood assay. Human whole blood secretes TNFxcex1 when stimulated with LPS. This property of blood forms the basis of an assay which is used as a secondary test for compounds which profile as active in the PBMC test.
Heparinised (10 units/ml) human blood was obtained from volunteers. 160 xcexcl whole blood were added to 96 well round-bottom plates (Corning 25850). Compounds were solubilised and serially diluted in RPMI 1640 medium (Gibco) supplemented with 50 units/ml penicillin, 50 xcexcg/ml streptomycin and 2 mM glutamine, as detailed above. 20 xcexcl of each test concentration was added to appropriate wells (triplicate cultures). 20 xcexcl of RPMI 1640 medium supplemented with antibiotics and glutamine was added to control wells. Plates were incubated for 30 minutes at 37xc2x0 C. (humidified incubator), prior to addition of 20 xcexcl LPS (final concentration 10 xcexcg/ml). RPMI 1640 medium was added to control wells. Six xe2x80x9cLPS alonexe2x80x9d and four xe2x80x9cmedium alonexe2x80x9d controls were included on each plate. A known TNFxcex1 synthesis/secretion inhibitor was included in each test. Plates were incubated for 6 hours at 37xc2x0 C. (humidified incubator). Plates were centrifuged (2000 rpm for 10 minutes) and 100 xcexcl plasma removed and stored at xe2x88x9270xc2x0 C. (Corning 25850 plates). TNFxcex1 levels were measured by ELISA (see WO92/10190 and Current Protocols in Molecular Biology, vol 2 by Frederick M. Ausbel et al., John Wiley and Sons Inc.). The paired antibodies that were used in the ELIZA were obtained from RandD Systems (catalogue nos. MAB610 anti-human TNFxcex1 coating antibody, BAF210 biotinylated anti-human TNFxcex1 detect antibody).
Ex Vivo/In Vivo Assessment
The ability of the compounds of this invention as ex vivo TNFxcex1 inhibitors were assessed in the rat or mouse. Briefly, groups of male Wistar Alderley Park (AP) rats (180-210 g) were dosed with compound (6 rats) or drug vehicle (10 rats) by the appropriate route, for example peroral (p.o.), intraperitoneal(i.p.) or subcutaneous (s.c.). Ninety minutes later rats were sacrificed using a rising concentration of CO2 and bled out via the posterior vena cavae into 5 Units of sodium heparin/ml blood. Blood samples were immediately placed on ice and centrifuged at 2000 rpm for 10 min at 4xc2x0 C. and the harvested plasmas frozen at xe2x88x9220xc2x0 C. for subsequent assay of their effect on TNFxcex1 production by LPS-stimulated human blood. The rat plasma samples were thawed and 175 xcexcl of each sample was added to a set format pattern in a 96 well round bottom plate (Coming 25850). 50 xcexcl of heparinized human blood was then added to each well, mixed and the plate was incubated for 30 min at 37xc2x0 C. (humidified incubator). LPS (25 xcexcl; final concentration 10 xcexcg/ml) was added to the wells and incubation continued for a further 5.5 hours. Control wells were incubated with 25 xcexcl of medium alone. Plates were then centrifuged for 10 min at 2000 rpm and 200 xcexcl of the supernatants were transferred to a 96 well plate and frozen at xe2x88x9220xc2x0 C. for subsequent analysis of TNF concentration by ELISA.
Data analysis by dedicated software calculates for each compound/dose:       %    ⁢          xe2x80x83        ⁢    inhibition    ⁢          xe2x80x83        ⁢    of    ⁢          xe2x80x83        ⁢    TNF    ⁢          xe2x80x83        ⁢    α    =                              Mean          ⁢                      xe2x80x83                    ⁢          TNF          ⁢                      xe2x80x83                    ⁢                      α            ⁡                          (              Controls              )                                      -                  Mean          ⁢                      xe2x80x83                    ⁢          TNF          ⁢                      xe2x80x83                    ⁢                      α            ⁡                          (              Treated              )                                                  Mean        ⁢                  xe2x80x83                ⁢        TNF        ⁢                  xe2x80x83                ⁢                  α          ⁡                      (            Controls            )                                xc3x97    100  
Alternatively, mice could be used instead of rats in the above procedure.
Test as Anti-Arthritic Agent
Activity of a compound as an anti-arthritic agent was tested as follows. Acid soluble native type II collagen was shown by Trentham et al. [1] to be arthritogenic in rats; it caused polyarthritis when administered in Freunds incomplete adjuvant. This is now known as collagen-induced arthritis (CIA) and similar conditions can be induced in mice and primates. Recent studies have shown that anti-TNF monoclonal antibodies [2] and TNF receptor-IgG fusion proteins [3] ameliorate established CIA indicating that TNF plays a key role in the pathophysiology of CIA. Moreover, the remarkable efficacy reported for anti-TNF monoclonal antibodies in recent rheumatoid arthritis clinical trials indicates that TNF plays a major role in this chronic inflammatory disease. Thus CIA in DBA/1 mice as described in references 2 and 3 is a tertiary model which can be used to demonstrate the anti-arthritic activity of a compound. Also see reference 4.
1. Trentham, D. E. et al., (1977) J. Exp. Med., 146 857.
2. Williams, R. O. et al., (1992) Proc. Natl. Acad. Sci. 89, 9784.
3. Williams, R. O. et al., (1995) Immunology, 8, 433.
4 Badger, M. B. et al., (1996) The Journal of Pharmacology and Experimental Therapeutics, 279, 1453-1461.
Although the pharmacological properties of the compounds of the Formula (I) vary with structural change as expected, in general a compound of the Formula (I) gives over 30% inhibition of p38xcex1 and/or p38xcex2 at concentrations up to 10 xcexcM and over 30% inhibition in the PBMC test at concentrations up to 50 xcexcM. No physiologically unacceptable toxicity was observed at the effective dose for compounds tested of the present invention. By way of example:xe2x80x94